Fig. 2: Autophagic vescicles accumulation upon RTN-1C induction.

a–g Ultrastructural analyses of SH-SY5Y neuroblastoma controls cells (a, b) or overexpressing RTN1-C for 24 h (c–g). N nucleus, m mitochondria, AV autophagic vesicles. Scale bars: 1 μm. h The number of autophagic vacuoles were counted under the Zeiss EM 900 electron microscope at 12.000x magnification (48 μm²) for each treatment conditions. Autophagic vacuoles were classified as autophagosomes when met two or more of the following criteria: double membrane, compartments of 0.5 μm in diameter or larger, luminal uncompacted cytosolic material including organelles, absence of ribosomes attached to the cytosolic side of the membrane. Were examined 70–100 fields per treatment condition and value are expressed as AVs per field. Finally, data were averaged to median values ± standard deviation (±SD) and used for statistical analysis. (***) (P < 0.001 vs. Ctrl group, Student’s t test).